Discovery of novel inhibitors of human S-adenosylmethionine decarboxylase based on in silico high-throughput screening and a non-radioactive enzymatic assay.

Chenzeng Liao,Yanlin Wang,Xiao Tan,Lidan Sun,Sen Liu

doi:10.1038/srep10754

Chenzeng Liao, Yanlin Wang + Show 3 more

Open Access

https://doi.org/10.1038/srep10754

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Journal: Scientific Reports	Publication Date: Jun 1, 2015
Citations: 15	License type: CC BY 4.0

Affiliation: China Three Gorges University

Abstract

Natural polyamines are small polycationic molecules essential for cell growth and development, and elevated level of polyamines is positively correlated with various cancers. As a rate-limiting enzyme of the polyamine biosynthetic pathway, S-adenosylmethionine decarboxylase (AdoMetDC) has been an attractive drug target. In this report, we present the discovery of novel human AdoMetDC (hAdoMetDC) inhibitors by coupling computational and experimental tools. We constructed a reasonable computational structure model of hAdoMetDC that is compatible with general protocols for high-throughput drug screening, and used this model in in silico screening of hAdoMetDC inhibitors against a large compound library using a battery of computational tools. We also established and validated a simple, economic, and non-radioactive enzymatic assay, which can be adapted for experimental high-throughput screening of hAdoMetDC inhibitors. Finally, we obtained an hAdoMetDC inhibitor lead with a novel scaffold. This study provides both new tools and a new lead for the developing of novel hAdoMetDC inhibitors.

Highlights

Background checking assayTo check the background effects of inhibitors, different concentrations of inhibitors were added to the reaction system similar as the enzymatic activity assay, except that hAdoMetDC was not included, or the NaHCO3 standard was used
The Phosphoenolpyruvate Decarboxylase (PEPC)-malate dehydrogenase (MDH) method was widely used for detecting CO2 produced by decarboxylases (ODC, for example)[1,19,20,21,32,34,35,36,37,38,39]
The first step, catalyzed by Phosphoenolpyruvate Decarboxylase (PEPC), is the bicarbonate condenses with phosphoenol pyruvate to form oxalate, and in the second step, oxalate is enzymatically reduced by Malate Dehydrogenase (MDH, using an NADH cofactor) to form malate and NAD+

Summary

Introduction

Background checking assayTo check the background effects of inhibitors (compounds), different concentrations of inhibitors (compounds) were added to the reaction system similar as the enzymatic activity assay, except that hAdoMetDC was not included, or the NaHCO3 standard was used. We report the screening of a novel hAdoMetDC inhibitor lead by integrated computational and experimental HTP assays.

Results

Conclusion